CA2249212A1 - Methods of tissue induction using a combination of bone morphogenetic protein and parathyroid hormone-related peptide - Google Patents
Methods of tissue induction using a combination of bone morphogenetic protein and parathyroid hormone-related peptide Download PDFInfo
- Publication number
- CA2249212A1 CA2249212A1 CA002249212A CA2249212A CA2249212A1 CA 2249212 A1 CA2249212 A1 CA 2249212A1 CA 002249212 A CA002249212 A CA 002249212A CA 2249212 A CA2249212 A CA 2249212A CA 2249212 A1 CA2249212 A1 CA 2249212A1
- Authority
- CA
- Canada
- Prior art keywords
- bmp
- composition
- proteins
- pthrp
- compositions
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/29—Parathyroid hormone (parathormone); Parathyroid hormone-related peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/04—Drugs for skeletal disorders for non-specific disorders of the connective tissue
Abstract
Compositions of proteins with chondrocyte and cartilaginous tissue inducing activity, as well as method of using those compositions, are disclosed. The compositions comprise one or more proteins of the transforming growth factor-.beta. (TGF-.beta.) superfamily of proteins, particularly bone morphogenetic proteins (BMPs), in combination with parathyroid hormone-related polypeptide (PTHrP) or an equivalent PTH-like polypeptide. The compositions and methods are useful in the treatment of osteoarthritis, cartilage defects and in related tissue repair.
Description
wo 97135607 PCT/USg7/04162 TITLE OF THE INVENTION
METHODS OF TISSUE INDUCTION USING A COMBINATION OF
BONE MORPHOGENETIC PROTEIN AND PARATHYROID HORMONE-RELATED I ~l~ ll)E
FIELD OF THE INVENTION
The present invention relates to novel methods and compositions for repairing, reducing or preventing damage to cartilage and cartilaginous tissue. The methods and compositions may further be useful for the induction and maintenanceof cartilaginous tissue formation, wound healing and cartilage and other tissue repair.
These methods and compositions may also be useful for ~ugm~onting the activity of bone morphogenetic proteins. In particular, the present application relates to the use of compositions comprising an osteogenic or cartilage-inducing member of the transforming factor beta [TGF-,B] superfamily of proteins, such as a bone morphogenetic protein [BMP], in combination with parathyroid hotmone-related peptide [PTHrP~. The compositions are useful for induction and maintenance of cartilaginous tissue, such as articular cartilage.
BACKGROUND OF THE INVENTION
The search for the molecule or molecules responsible for formation of bone, cartilage, tendon and other tissues present in bone and other tissue extracts has led to the discovery of a novel set of molecules called the Bone Morphogenetic Proteins (BMPs). The structures of several proteins, ~esign~te~l BMP-1 through BMP-15, have previously been elucidated. The unique inductive activities of these proteins, along with their presence in bone, cartilage and/or other vital tissues, suggests that they are important regulators of bone and other tissue repair processes, and may be involved in tissue formation, m~int~n~nre and repair. There is a need to identify improved methods and compositions for formation, maintenance and repair of such tissues.
Members of the bone morphogenetic protein family have been shown to be useful for induction of cartilage and bone formation. For example, BMP-2 has been shown to be able to induce the formation of new cartilage and/or bone tissue in vivo S(l~STITUTE SHEET (RULE 26) W O 97/35607 PCTrUS97/04162 in a rat ectopic implant model, see United States Patent No. 5,013,649; in mandibular defects ;n dogs, see Toriumi et al., Arch. Otolaryn~ol Head Neck Sur~., 117:1101-1112 ~1991); in femoral segm~nt~l defects in sheep, see Gerhart et al.,Trans Orthop Res Soc, 16: 172 (1991). Other members of the BMP family have also 5 been shown to have osteogenic activity, including BMP-4, -6 and -7, see Wozney, Bone Mo~phogenetic Proteins and Their Gene Erpression, in Cellular and MolecularBiology of Bone, pp. 131-167 (Academic Press, Inc. 1993). BMP proteins have alsobeen shown to demonstrate inductive and/or differentiation potentiating activity on a variety of other tissues, including cartilage, tendon, ligament, neural tissue.
Parathyroid hormone-related peptide (PTHrP) is a protein which is known exist in at least three isoforms of 139, 141 and 173 amino acids. [Karaplis et al., Genes & Development, 8:277-289 (1994). PTHrP is highly homologous to the N-terminal fragment of parathyroid hormone (PTH), and binds the same receptor as PTH. PTHrP appears to play a substantial role in calcium metabolism by an 15 autocrine/paracrine mech~ni~m, and also appears to regulate embryonal development, vascular tone and nutrition. Ts-lk~7~ki et al., Calcif Tissue Int 57: 196-200 (1995).
It has recently been shown that mice deficient in PTHrP exhibit abnormal cartilage maturation, in-ii( ating that PTHrP is an essential factor for chondrocyte development and Jllal~lraLion. In Ts -k~7.~ki, it is reported that PTHrP expression in articular 20 cartilage varies in hlLensiLy and loc~li7~tion during development, while the PTH/PTHrP receptor is highly expressed in the growth plate and in articular cartilage.
SUMMARY OF THE ~VENTION
The present invention relates to compositions useful for inducing cartilaginous 25 tissue formation in a patient in need of same, said compositions comprising one or more protein members of the transforming growth factor-~ (TGF-O superfamily together with parathyroid hormone related peptide (PTHrP), or an equivalent PTH-like polypeptide. Preferably, the compositions of the invention comprise at least one protein member of the bone morphogenetic protein (BMP) family and parathyroid 30 hormone related peptide (PTHrP). In particular plefelled embodiments, the composition colllplises a bone morphogenetic protein is selected from the group con~ictin~ of BMP-2, BMP-4, BMP-5, BMP-7, BMP-12 and BMP-13 together with SVBSTITUTE SHEET (RULE 26) wo 9713s607 PCT/US97/04162 PTHrP or an equivalent PTH-like polypeptide. In the most preferred embodiments, the compositions comprise BMP-2, BMP-13, heterodimers of BMP-2 and BMP-13 or combinations of the above, and PTHrP or an equivalent PTH-like polypeptide;
especially preferred BMPs are BMP-2 and BMP-13. The compositions may further 5 comprise one or more additional members of the BMP subfamily of proteins. In preferred embodiments of the invention, the PTHrP used is PTHrP1-34, a truncatedpeptide comprising the first 34 amino acids of the N-terminal portion of PTH. Inpl~fe.red embodiments of the invention, the compositions comprise BMP-2 and one or more additional proteins selected from the group consisting of BMP-4, BMP-5, BMP-7, BMP-12 and BMP-13; or BMP-13 and one or more additional proteins selected from the group consisting of BMP-2, BMP-4, BMP-S, BMP-7 and BMP-12 together with PTHrP1-34, PTHrP or another PTH-like polypeptide.
In other embodiments, the present invention relates to methods for inducing the formation and m~inten:lnre of cartilage in a patient, for example a patient 15 suffering from arthritis, particularly osteoarthritis, or a patient with an articular cartilage defect or other cartilaginous tissue defect, said method COlllp~ isillg a~1mini.~tering to said patient an effective amount of the above compositions. In a particular embodiment, the method of the present invention relates to a method for treating articular cartilage defects or damage in a patient in need of same, said 20 method comprising a~mini~t-oring to said patient an effective amount of the above compositions. The invention further relates to methods for inducing the formation of cartilage and cartilaginous tissue comprising a~lmini~tering to a patient a composition comprising a member of the TGF-,B ~ul,tlra...ily of p-otehls and Parathyroid hormone-related peptide (PTHrP).
The methods and compositions of the present invention are thus useful for repairing, reducing or preventing damage to cartilage and cartilaginous tissue. The methods and compositions may further be useful for the induction and mainlenallce of cartilaginous tissue formation, wound healing and cartilage and other tissue repair.
for the induction of cartilaginous tissue, such as articular cartilage, the meni~c~
and the articular surfaces of developing bone, or for the treatment of diseases or defects of cartilaginous tissue, such as arthritis, particularly osteoarthritis.
SUBS~ITUTE SHEET (RULE 26) W O 97135607 PCTrUS97/04162 DETAILED DESCRIPIION OF THE INVENTION
The methods and compositions of the present invention comprise a combination of one or more proteins from the transforming growth factor-~
superfamily with one or more parathyroid hormone-related peptides. The S transforming growth factor-,~ superfamily is a well-characterized family of proteins involved in cellular proliferation and dirf~rc,lliation of cells into various tissues.
Members of the TGF-,l~ superfamily are generally dimeric in structure, comprising two monomeric units which are produced by proteolytic cleavage from a larger precursor protein, of which the processed monomer represents the carboxyl terminal portion. The dimeric TGF-,~ proteins generally have molecular weights of approximately 20,000 to 35,000 and share a common cysteine pattern in the matureprotein region. See, for example, Sporn et al ., Science, 233 :532-534 (1986) and the papers cited therein. The TGF-~ superfamily includes several subgroups beside TGF-~1 through -,BS, are the bone morphogenetic proteins (BMPs), growth and differentiation factors (GDFs), the inhibins, as well as GDNF and Mullerian inhibitory substance and other structurally related proteins. The TGF-,B superfamily also includes proteins from other species, which have been characterized and arehighly conserved compared to the m~mm~ n TGF-,l~s, including Vgl (Xenopus), see for e~cample, Weeks and Melton, Cell, 51 :861 -867 (1987); Dpp, Screw and 60A
(Drosophila), see for example Padgett et al., Nah~re (London), 325:81-84 (1987);Doctor et al., Dev. Biol. 151:591-505 (1992); and more recently identified proteins including Univin (sea urchin), Dorsalin-1 (chicken) and Radar (Zebrafish). Otherfactors which may be effectively used in the composition include synthetic molecules or fragments of a TGF-~ supclrallJily member which are able to bind to a TGF-~
receptor molecule.
Methods for production of l,u,llcl.~us members of the TGF-,(~ superfamily useful in the present invention are known and/or described in the literature. For example, the structure and methods for production of many BMPs, including BMP-2,BMP-3, BMP-4, BMP-S, BMP-6 and BMP-7, are disclosed, for inct~nre, in United States Patents 5,108,922; 5,013,649; 5,116,738; 5,106,748; 5,187,076; and 5,141,905; BMP-8, disclosed in PCT publication WO91118098; BMP-9, disclosed in PCT publication W093/00432; BMP-10, disclosed in PCT application SUBSTITUTE SHEET (RULE 26) W 097/35607 PCT~US97/04162 W094/26893; BMP-11, disclosed in PCT application W094/26892; BMP-12 and BMP-13, disclosed in PCT application WO 95!16035, or BMP-15, disclosed in co-pending patent application, serial no. 08/446,924, filed on May 18, 1995. The structure of Vgr-2, and the growth and dirr~le.llidLion factors ~GDFs], including S those described in PCT applications WO94/15965; WO94/15949; WO95/01801;
METHODS OF TISSUE INDUCTION USING A COMBINATION OF
BONE MORPHOGENETIC PROTEIN AND PARATHYROID HORMONE-RELATED I ~l~ ll)E
FIELD OF THE INVENTION
The present invention relates to novel methods and compositions for repairing, reducing or preventing damage to cartilage and cartilaginous tissue. The methods and compositions may further be useful for the induction and maintenanceof cartilaginous tissue formation, wound healing and cartilage and other tissue repair.
These methods and compositions may also be useful for ~ugm~onting the activity of bone morphogenetic proteins. In particular, the present application relates to the use of compositions comprising an osteogenic or cartilage-inducing member of the transforming factor beta [TGF-,B] superfamily of proteins, such as a bone morphogenetic protein [BMP], in combination with parathyroid hotmone-related peptide [PTHrP~. The compositions are useful for induction and maintenance of cartilaginous tissue, such as articular cartilage.
BACKGROUND OF THE INVENTION
The search for the molecule or molecules responsible for formation of bone, cartilage, tendon and other tissues present in bone and other tissue extracts has led to the discovery of a novel set of molecules called the Bone Morphogenetic Proteins (BMPs). The structures of several proteins, ~esign~te~l BMP-1 through BMP-15, have previously been elucidated. The unique inductive activities of these proteins, along with their presence in bone, cartilage and/or other vital tissues, suggests that they are important regulators of bone and other tissue repair processes, and may be involved in tissue formation, m~int~n~nre and repair. There is a need to identify improved methods and compositions for formation, maintenance and repair of such tissues.
Members of the bone morphogenetic protein family have been shown to be useful for induction of cartilage and bone formation. For example, BMP-2 has been shown to be able to induce the formation of new cartilage and/or bone tissue in vivo S(l~STITUTE SHEET (RULE 26) W O 97/35607 PCTrUS97/04162 in a rat ectopic implant model, see United States Patent No. 5,013,649; in mandibular defects ;n dogs, see Toriumi et al., Arch. Otolaryn~ol Head Neck Sur~., 117:1101-1112 ~1991); in femoral segm~nt~l defects in sheep, see Gerhart et al.,Trans Orthop Res Soc, 16: 172 (1991). Other members of the BMP family have also 5 been shown to have osteogenic activity, including BMP-4, -6 and -7, see Wozney, Bone Mo~phogenetic Proteins and Their Gene Erpression, in Cellular and MolecularBiology of Bone, pp. 131-167 (Academic Press, Inc. 1993). BMP proteins have alsobeen shown to demonstrate inductive and/or differentiation potentiating activity on a variety of other tissues, including cartilage, tendon, ligament, neural tissue.
Parathyroid hormone-related peptide (PTHrP) is a protein which is known exist in at least three isoforms of 139, 141 and 173 amino acids. [Karaplis et al., Genes & Development, 8:277-289 (1994). PTHrP is highly homologous to the N-terminal fragment of parathyroid hormone (PTH), and binds the same receptor as PTH. PTHrP appears to play a substantial role in calcium metabolism by an 15 autocrine/paracrine mech~ni~m, and also appears to regulate embryonal development, vascular tone and nutrition. Ts-lk~7~ki et al., Calcif Tissue Int 57: 196-200 (1995).
It has recently been shown that mice deficient in PTHrP exhibit abnormal cartilage maturation, in-ii( ating that PTHrP is an essential factor for chondrocyte development and Jllal~lraLion. In Ts -k~7.~ki, it is reported that PTHrP expression in articular 20 cartilage varies in hlLensiLy and loc~li7~tion during development, while the PTH/PTHrP receptor is highly expressed in the growth plate and in articular cartilage.
SUMMARY OF THE ~VENTION
The present invention relates to compositions useful for inducing cartilaginous 25 tissue formation in a patient in need of same, said compositions comprising one or more protein members of the transforming growth factor-~ (TGF-O superfamily together with parathyroid hormone related peptide (PTHrP), or an equivalent PTH-like polypeptide. Preferably, the compositions of the invention comprise at least one protein member of the bone morphogenetic protein (BMP) family and parathyroid 30 hormone related peptide (PTHrP). In particular plefelled embodiments, the composition colllplises a bone morphogenetic protein is selected from the group con~ictin~ of BMP-2, BMP-4, BMP-5, BMP-7, BMP-12 and BMP-13 together with SVBSTITUTE SHEET (RULE 26) wo 9713s607 PCT/US97/04162 PTHrP or an equivalent PTH-like polypeptide. In the most preferred embodiments, the compositions comprise BMP-2, BMP-13, heterodimers of BMP-2 and BMP-13 or combinations of the above, and PTHrP or an equivalent PTH-like polypeptide;
especially preferred BMPs are BMP-2 and BMP-13. The compositions may further 5 comprise one or more additional members of the BMP subfamily of proteins. In preferred embodiments of the invention, the PTHrP used is PTHrP1-34, a truncatedpeptide comprising the first 34 amino acids of the N-terminal portion of PTH. Inpl~fe.red embodiments of the invention, the compositions comprise BMP-2 and one or more additional proteins selected from the group consisting of BMP-4, BMP-5, BMP-7, BMP-12 and BMP-13; or BMP-13 and one or more additional proteins selected from the group consisting of BMP-2, BMP-4, BMP-S, BMP-7 and BMP-12 together with PTHrP1-34, PTHrP or another PTH-like polypeptide.
In other embodiments, the present invention relates to methods for inducing the formation and m~inten:lnre of cartilage in a patient, for example a patient 15 suffering from arthritis, particularly osteoarthritis, or a patient with an articular cartilage defect or other cartilaginous tissue defect, said method COlllp~ isillg a~1mini.~tering to said patient an effective amount of the above compositions. In a particular embodiment, the method of the present invention relates to a method for treating articular cartilage defects or damage in a patient in need of same, said 20 method comprising a~mini~t-oring to said patient an effective amount of the above compositions. The invention further relates to methods for inducing the formation of cartilage and cartilaginous tissue comprising a~lmini~tering to a patient a composition comprising a member of the TGF-,B ~ul,tlra...ily of p-otehls and Parathyroid hormone-related peptide (PTHrP).
The methods and compositions of the present invention are thus useful for repairing, reducing or preventing damage to cartilage and cartilaginous tissue. The methods and compositions may further be useful for the induction and mainlenallce of cartilaginous tissue formation, wound healing and cartilage and other tissue repair.
for the induction of cartilaginous tissue, such as articular cartilage, the meni~c~
and the articular surfaces of developing bone, or for the treatment of diseases or defects of cartilaginous tissue, such as arthritis, particularly osteoarthritis.
SUBS~ITUTE SHEET (RULE 26) W O 97135607 PCTrUS97/04162 DETAILED DESCRIPIION OF THE INVENTION
The methods and compositions of the present invention comprise a combination of one or more proteins from the transforming growth factor-~
superfamily with one or more parathyroid hormone-related peptides. The S transforming growth factor-,~ superfamily is a well-characterized family of proteins involved in cellular proliferation and dirf~rc,lliation of cells into various tissues.
Members of the TGF-,l~ superfamily are generally dimeric in structure, comprising two monomeric units which are produced by proteolytic cleavage from a larger precursor protein, of which the processed monomer represents the carboxyl terminal portion. The dimeric TGF-,~ proteins generally have molecular weights of approximately 20,000 to 35,000 and share a common cysteine pattern in the matureprotein region. See, for example, Sporn et al ., Science, 233 :532-534 (1986) and the papers cited therein. The TGF-~ superfamily includes several subgroups beside TGF-~1 through -,BS, are the bone morphogenetic proteins (BMPs), growth and differentiation factors (GDFs), the inhibins, as well as GDNF and Mullerian inhibitory substance and other structurally related proteins. The TGF-,B superfamily also includes proteins from other species, which have been characterized and arehighly conserved compared to the m~mm~ n TGF-,l~s, including Vgl (Xenopus), see for e~cample, Weeks and Melton, Cell, 51 :861 -867 (1987); Dpp, Screw and 60A
(Drosophila), see for example Padgett et al., Nah~re (London), 325:81-84 (1987);Doctor et al., Dev. Biol. 151:591-505 (1992); and more recently identified proteins including Univin (sea urchin), Dorsalin-1 (chicken) and Radar (Zebrafish). Otherfactors which may be effectively used in the composition include synthetic molecules or fragments of a TGF-~ supclrallJily member which are able to bind to a TGF-~
receptor molecule.
Methods for production of l,u,llcl.~us members of the TGF-,(~ superfamily useful in the present invention are known and/or described in the literature. For example, the structure and methods for production of many BMPs, including BMP-2,BMP-3, BMP-4, BMP-S, BMP-6 and BMP-7, are disclosed, for inct~nre, in United States Patents 5,108,922; 5,013,649; 5,116,738; 5,106,748; 5,187,076; and 5,141,905; BMP-8, disclosed in PCT publication WO91118098; BMP-9, disclosed in PCT publication W093/00432; BMP-10, disclosed in PCT application SUBSTITUTE SHEET (RULE 26) W 097/35607 PCT~US97/04162 W094/26893; BMP-11, disclosed in PCT application W094/26892; BMP-12 and BMP-13, disclosed in PCT application WO 95!16035, or BMP-15, disclosed in co-pending patent application, serial no. 08/446,924, filed on May 18, 1995. The structure of Vgr-2, and the growth and dirr~le.llidLion factors ~GDFs], including S those described in PCT applications WO94/15965; WO94/15949; WO95/01801;
2; W094/21681; WO94/15966; and others are also known. Other TGF-~ proteins which may be useful in the present invention include BIP, disclosed in WO94/01557; and MP52, disclosed in PCT application WO93/16099. Methods for production of heterodimeric proteins comprising two distinct monomeric units, each co.ll~lisillg the amino acid sequence of one of the above TGF-~ proteins, are described in W093/09229. The disclosures of all of the above applications are hereby incorporated by reference.
Parathyroid hormone-related peptide (PTHrP) is closely related in structure to parathyroid hormone. PTHrP is known to exist in at least three isoforms of 139, 141 and 173 amino acids. [Karaplis et al., Genes & Development, 8:277-289 (1994).
PTHrP is highly homologous to the N-terminal fragment of parathyroid hormone (PTH), and binds the same receptor as PTH. The nucleotide and amino acid sequences of the PTHrP gene from rat, mouse and human are known and may be used to produce PTHrP-like polypeptides useful in the present invention. [See Karaplis et al., Mol. Endocrin. 4:441-446 [1990][rat]; Mangin et al., PNAS
85 :597-601 (1988)[human] and Mangin et al., Gene 95:195-202 (1990)[mouse]; and Martin et al., Crit Rev Biochem Mal Biol 26:377-395 (1991).] In a preferred embodiment of the present invention, a variant of PTHrP is used in which one or more amino acids from the carboxy terminus has been deleted. For example, PTHrPl-34, which comprises the first 34 amino acids of PTHrP, is used in one preferred embodiment of the present invention. Also useful in the present invention are PTH-like polypeptides which are equivalent to PTHrPl-34 in their ability to enhance survival of chondrocytes. Such PTH-like polypeptides may include, for example, PTH, whether of human, porcine, bovine or other ~ n origin;
variants of PTH, such as those described in Wingender et al., United States Patent 5,455,329; Wingender et al., Unioed States Patent 5,457,047; and Schluter et al., United States Patent 5,457,092, and the references cited therein; as well as variants SUBSTITUTE SHEET (RULE 26) wo 97/35607 PCT/US97/04162 of the above in which one or more amino acids of PTH has been deleted from the carboxy andlor amino tennin~l portions of the molecule. The disclosures of the above publications are hereby incorporated by reference. PTH, PTHrP and the above variants may be produced via recombinant DNA engineering using the known 5 sequences of the PTH and PTHrP proteins, or may be isolated by purification.
The methods and compositions of the present invention may comprise simultaneous or sequential ~-lmini~tration of at least two active agents, a TGF~protein and a parathyroid hormone-related peptide, to a patient or site in need of cartilage repair, formation or maintenance. For sequential a~lmini~tration, one or 10 both of the active agents may be encapsulated or otherwise m~int~in~d in contact with a carrier which provides for slow release of the agent. The methods and compositions may comprise the active agents in a weight ratio of from about 90:10, to about 10:90. Preferably the active agents are present in a weight ratio of about 70:30 to 30:70. Most preferred are methods and compositions co~ lisillg the active 15 agents in a weight ratio of about 50:50.
The compositions of the invention may comprise, in addition to a TGF-,B
protein and a parathyroid hormone-related peptide, other therapentic~lly useful agents including growth factors such as epidermal growth factor (EGF), transforming growth factor-c~, activins, inhibins, platelet derived growth factor (PDGF), fibroblast 20 growth factor (FGF), and fibroblast growth factor-4 (FGF-4), parathyroid hormone (PTH), lellkt~rni~ inhibitory factor (LIF/HILDA/DIA), and insulin-like growth factors (IGF-I and IGF-II). Portions of these agents may also be used in compositions ofthe present invention. The compositions may also include an appl~l3liate matrix for in~t~nre, for supporting the composition and providing a surface for cartilage or for 25 other connective tissue growth. The matrix may provide slow release of the protein and/or the appr~liate environment for presentation thereof.
The methods and compositions of the present invention employ proteins which are able to induce cartilaginous tissue or other tissue formation in circl~Tn~t~nres where such tissue is not normally formed, and has application in the healing of 30 cartilage, for example articular cartilage tears, deformities and other cartilage defects in hllm~n~ and other animals. Such methods and compositions employing cartilaginous tissue inducing proteins may have prophylactic use in preventing SUBSTITUTE SHEET (RULE 26) damage to cartilaginous tissue, as well as use in the improved fixation of cartilage to bone or other tissues, and in repairing defects to cartilage tissue. De novo cartilaginous tissue formation induced by a composition of the present inventioncontributes to the repair of congenital, trauma in(ll~ced, or other cartilage defects of 5 other origin, and is also useful in surgery for attachment or repair of cartilage. The methods and compositions of the invention may also be useful in the treatment ofarthritis and other cartilage defects. The methods and compositions of the present invention can also be used in other indications wherein it is desirable to heal or regenerate cartilage tissue. Such indications include, without limitation, regeneration 10 or repair of injuries to the articular cartilage. The methods and compositions of the present invention may provide an environment to attract cartilage-forming cells,stim~ te growth of cartilage-forming cells or induce differentiation of progenitors of cartilage-forming cells and chondrocytes.
The compositions and methods of the present invention may also be useful 15 for treating cell populations, such as embryonic cells or stem cell populations, to enhance or enrich the growth, survival and/or differentiation of the cells into chondrocytes or other cell types. In another embodim~nt, the compositions and methods of the present invention may be used to treat chondrocytic cell lines, such as articular chondrocytes, in order to m~int~in chondrocytic phenotype and survival 20 of the cells. The treated cell populations may be useful for gene therapy applications.
The proteins useful in the methods of the present invention are useful for inducing the formation, m~ te~ re and survival of chondrocytes and/or cartilaginous tissue. It is contemplated that these proteins may have the ability to 25 induce the formation of other types of tissue, such as tendon and lig~mPnt, as well.
The cartilaginous tissue-inducing methods and compositions provided herein also may include factors encoded by the sequences similar to those of naturally-occurringTGF-,B l~lo~eins, into which modifications are naturally provided (e.g. allelic variations in the nucleotide seq-lenre which may result in amino acid changes in the 30 polypeptide) or deliberately engineered. For example, synthetic polypeptides may wholly or partially duplicate continuous sequences of the amino acid residues ofBMP-2 or BMP-13. These sequences, by virtue of sharing primary, secondary, or SUBSrlTUTE SHEET (RUL~ 26) W O 97/356D7 PCT~US97/04162 tertiary structural and conformational characteristics with cartilaginous tissue growth factor polypeptides of naturally-occurring BMP-2 or BMP-13 may possess cartilaginous or other tissue growth factor biological properties in comrnon thelcwilh. Thus, they may be employed as biologically active substitutes for 5 naturally-occurring cartilaginous tissue inducing polypeptides in therapeutic methods and compositions.
The proteins useful in the present invention, or the DNA sequences encoding therefor, may be engineered to provide one or more additional cysteine residues to increase potential dimer formation, particularly for forming dimers of the TGF-~
10 proteins. The resulting DNA sequence would be capable of producing a "cysteineadded variant" of the protein. Production of "cysteine added variants" of proteins is described in United States Patent 5,166,322, the disclosure of which is hereby incorporated by reference. Other specific mutations of the sequences of cartilaginous tissue inducing l.roteills described herein involve modifications of glycosylation sites.
15 These modifications may involve O-linked or N-linked glycosylation sites. Forin~t~nce, the absence of glycosylation or only partial glycosylation results from amino acid substitution or deletion at asparagine-linked glycosylation recognition sites. The asparagine-linked glycosylation recognition sites comprise tripeptidesequences which are specifically recognized by apprcpliate cellular glycosylation 20 enzymes. These tripeptide sequenre~ may be asparagine-X-threonine, asparagine-X-serine or asparagine-X-cysteine, where X is usually any amino acid except proline.
A variety of amino acid substitutions or deletions at one or both of the first or third amino acid positions of a glycosylation recognition site (and/or amino acid deletion at the second position) results in non-glycosylation at the modified tripeptide 25 sequence. Additionally, bacterial expression of protein will also result in production of a non-glycosylated protein, even if the glycosylation sites are left unrnodified.
The methods of the present invention for inducing cartilaginous tissue forrnation may comprise a~lministering to a patient an effective amount of a composition colll~ hlg one or more protein members of the transforming growth 30 factor-,~ (TGF-,B) ~up~lralnily together with parathyroid hormone related peptide (PTHrP), or an equivalent PTH-like polypeptide. Alternatively, the methods of the present invention for inducing cartilaginous tissue formation may comprise SU8STITUTE SHEE~ (RULE 26) W 097/3S607 PCTrUS97/04162 heterodimers comprised of two distinct monomeric units, each of which has the amino acid sequence of a member of the TGF-,B superfamily, together with PTHrP, or an equivalent PTH-like polypeptide. Preferably, the compositions of the invention comprise one or more members of the bone morphogenetic protein (BMP) family, together with PTHrP or an equivalent PTH-like polypeptide. In particular preferred embodiments, the composition comprises a bone morphogenetic protein selected from the group con~i~ting of BMP-2,BMP-4,BMP-5,BMP-7,BMP-12 and BMP-13, or heterodimers of the above, together with PTHrP or an equivalent PTH-like polypeptide. In the most preferred embo~im~n~, the composition CO~ S BMP-2, BMP-13, heterodimers of BMP-2 and BMP-13 or combinations of the above, and PTHrP or an equivalent PTH-like polypeptide. The above compositions may optionally comprise one or more additional proteins which are members of the TGF-superfamily, preferably of the BMP family of proteins. In preferred embodiments,the composition comprises BMP-2 and one or more additional proteins selected from 15 the group consisting of BMP-4, BMP-5, BMP-7, BMP-12, BMP-13 and heterodimers of the above; or BMP-13 and one or more additional proteins selected from the group con.~i~ting of BMP-2,BMP-4,BMP-5,BMP-7,BMP-12 and heterodimers of the above; each together with PTHrP or an equivalent PTH-like polypeptide. As diccllcsed above, these compositions may be used to induce the 20 formation, mai.l~enallce and survival of chondrocytes and/or cartilaginous tissue or other tissue. It is contemplated that such compositions may also be used for articular cartilage repair, wound healing and other tissue repair, such as skin repair. It is further contemplated that proteins of the invention may increase n~uro~lal survival and therefore be useful in transplantation and treatment of conditions exhibiting a decrease in neuronal survival. Compositions and methods of the invention may further be useful for induction and repair of other tissue, including wound healing.
In a preferred embodiment of the present invention, more than one protein of the TGF-~ superfamily may be used together with PTHrP to enhance the induction, maintenance and diff~nliaLion of one or more cell and/or tissue types, 30 including cartilaginous tissue. For example, a composition comprising both BMP-2 and BMP-13 implanted together with PTHrP may give rise to both bone and cartilaginous tissue. Such a composition may be useful for treating defects of the SUBSTIJUTE SI~EET (RULE 26~
W O 97135607 PCTrUS97/04162 junction between cartilage and bone, causing cartilage and bone to form simultaneously at contiguous anatomical locations, and may therefore be useful for regenerating tissue at the site of cartilage attachment to bone.
It is contemplated that the methods and compositions of the invention may also be useful in wound healing, such as skin healing and related tissue repair. The types of wounds include, but are not limited to burns, incisions and ulcers. (See, e.g. PCT Publication WO84/01106 for discussion of wound healing and related tissue repair).
The methods and compositions of the invention further comprise heteromolecules comprised of different TGF-~ moieties, in combination with PTHrPor an equivalent PTH-like polypeptide. Preferably, the heteromolecule is a heterodimer, for example, of two monomers each c-,l"L,lisillg the amino acid sequence of a BMP protein. For example, a method and composition of the invention may comprise a ~ lfi-le linked dimer COIll~l iSillg a BMP-2 protein subunit and a subunit from one of the other BMP proteins described above. Thus, in a particular embo~iment, the present invention comprises compositions and methods employing a heterodimer wherein one subunit co,~lplises the amino acid sequence of BMP-2, and one subunit comprises an amino acid sequence for a bone morphogenetic protein selected from the group consisting of BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13 or BMP-15. Further embodiments may comprise a heterodimer of other disulfide bonded TGF-,B moieties, including VGR-2, MP52, MIS, activin and TGF-,~. For example, the heterodimer may comprise one subunit comprising the amino acid sequence of BMP-2 or BMP-13, and the other subunit may comprise the amino acid of activin.
Further, compositions of the present invention may be combined with other agentsbeneficial to the treatment of the defect, wound, or tissue in question.
It is expected that the compositions and methods of the invention may act in concert with or perhaps synergistically with ~lmini~tration of other related proteins and growth factors. Further therapeutic methods and compositions of the invention therefore comprise a thelapeulic amount of at least one of the BMP proteins, whether homodimeric or heterodimeric, in combination with PTHrP or an equivalent PTH-like polypeptide, and other growth factors.
SUBSTITUTE SHEET (RULE 26) wo 97/35607 PCT/US97/04162 The preparation and formulation of such physiologically acceptable protein compositions, having due regard to pH, isotonicity, stability and the like, is within the skill of the art. The therapeutic compositions are also presently valuable for veterinary applications due to the lack of species specificity in TGF-~ proteins.
5 Particularly domestic animals and thoroughbred horses in addition to humans are desired patients for such treatment with the compositions of the present invention.
The therapeutic method includes :~mini~tering the composition topically, systemically, or locally as an injectable and/or implant or device. When admin-istered, the therapeutic composition for use in this invention is, of course, in a 10 pyrogen-free, physiologically acceptable form. Further, the composition may desirably be encapsulated or injected in a viscous form for delivery to the site of tissue damage. Topical a(lmini~tration may be suitable for wound healing and tissue repair. Therapeutic~lly useful agents other than the proteins which may also optionally be included in the composition as described above, may alternatively or 15 additionally, be ~lmini~tered .siml~lt:~neously or sequentially with the composition in the methods of the invention. In addition, the compositions of the present invention may be used in conjunction with presently available treatments for cartilage injuries, such as suture (e.g., vicryl sutures or surgical gut sutures, Ethicon Inc., Somerville, NJ) or cartilage allograft or autograft, in order to enh~nre or accelerate 20 the healing potential of the suture or graft. For example, the suture, allograft or autograft may be soaked in the compositions of the present invention prior to implantation. It may also be possible to incorporate the protein or composition of the invention onto suture materials, for example, by freeze-drying.
The compositions of the present invention may include an app~ul)l iate matrix 25 and/or sequestering agent as a carrier. For in~t~nre, the matrix may support the composition or provide a surface for cartilaginous tissue formation and/or othertissue formation. The matrix may provide slow release of the protein and/or the ap~lopliate environment for presentation thereof. The sequestering agent may be a substance which aids in ease of a(lmini~tration through injection or other means, 30 or may slow the migration of protein from the site of application.
The choice of a carrier material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface SUBSTITIJTE SHEET (RULE 26) W 097/35607 PCT~US97/04162 properties. The particular application of the compositions will define the ~yplopfiate formulation. Potential matrices for the compositions may be biodegradable and chemically defined. Further matrices are comprised of pure proteins or extracellular matrix components. Other potential matrices are nonbiodegradable and chemically 5 defined. Preferred matrices include collagen-based materials, including sponges, such as Helistat~ (Integra LifeSciences, Plainsboro, N.J.), orcollagen in an injectable form, as well as sequestering agents, which may be biodegradable, for example hyaluronic acid derived. Biodegradable materials, such as cellulose films, or surgical meshes, may also serve as matrices. Such materials could be sutured into 10 an injury site, or wrapped around the cartilage.
Another preferred class of carrier are polymeric matrices, including polymers of poly(lactic acid), poly(glycolic acid) and copolymers of lactic acid and glycolic acid. These matrices may be in the form of a sponge, or in the form of porous particles, and may also include a sequçstering agent. Suitable polymer matrices are described, for example, in WO93/00050, the disclosure of which is incorporated herein by reference.
Preferred f~milil~s of se~uestering agents include blood, fibrin clot and/or cellulosic materials such as alkylcelluloses (including hydroxyalkylcelluloses),including methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl-methylcellulose, and carboxymethylcellulose, the most preferred being cationic salts of carboxymethylcellulose (CMC). Other preferred sequestering agents include hyaluronic acid, sodium alginate, poly(ethylene glycol), polyoxyethylene oxide, carboxyvinyl polymer and poly(vinyl alcohol). The amount of seql~estering agent useful herein is 0.5-20 wt%, preferably 1-10 wt%
based on total formulation weight, which represents the amount n~ces~ry to prevent desorbtion of the protein from the polymer matrix and to provide a~ opliate h~n-lling of the composition, yet not so much that the progenitor cells are prevented from infiltrating the matrix, thereby providing the protein the opportunity to assist the activity of the progenitor cells.
Additional optional components useful in the practice of the subject application include, e.g. cryogenic protectors such as m~nnitol, sucrose, lactose, glucose, or glycine (to protect the protein from degradation during Iyophilization), SUBSTITUTE SHEET (RULE 26) W O 97/35607 PCTrUS97/04162 antimicrobial preservatives such as methyl and propyl parabens and benzyl alcohol;
antioxidants such as EDTA, citrate and BHT (butylated hydroxytoluene); and surfactants such as poly(sorbates) and poly(oxyethylenes); etc.
As described above, the compositions of the invention may be employed in 5 methods for treating a number of cartilage defects, such as the regeneration of cartilaginous tissue in areas of cartilage damage, to assist in repair of tears of cartilage tissue, and various other types of tissue defects or wounds. These methods, according to the invention, entail ~Amini~tering to a patient nee(ling such cartilaginous tissue or other tissue repair, a composition comprising an effective 10 amount of an osteogenic or cartilage-inducing member of the transforming factor beta [TGF-~] superfamily of proteins, such as a bone morphogenetic protein [BMP], preferably BMP-2,BMP-4,BMP-S,BMP-7,BMP-12 and BMP-13, in combination with parathyroid hormone-related peptide [PTHrP] or an equivalent PTH-like polypeptide.
In another embodiment, the methods may entail ~lminictration of a heterodimeric protein in which each of the monomers cu~ lises the amino acid sequence of an osteogenic or cartilage-inducing member of the TGF,~ superfamily of proteins, such as a BMP, particularly, BMP-2,BMP-4,BMP-5,BMP-7,BMP-12 and BMP-13, in combination with PTHrP or an equivalent PTHrP-like polypeptide.
In a preferred embodiment, each of the monomers is selected from the group consisting of BMP-2,BMP-4,BMP-5,BMP-7,BMP-12 and BMP-13.
Thus, a further aspect of the invention is a therapeutic method and composition for inducing or m~int~ining chondrocytes or cartilaginous tissue, for repairing cartilaginous tissue, for repairing cartilage as well as treating arthritis and other conditions related to arthritis or cartilage defects. Such compositions comprise a therape~tir~lly effective amount of one or more osteogenic or cartilaginous tissue inducing proteins, such as BMP-2 or BMP-13, in combination with PTHrP or an equivalent PTH-like polypeptide, in admixture with a pharm~relltic~lly acceptable vehicle, carrier or matrix.
The dosage regimen will be det~rminrd by the attending physician considering various factors which modify the action of the composition, e.g., amount of cartilaginous tissue desired to be formed, the site of cartilaginous tissue damage, the SVBSrlTUTE SHEET (RUlE 26) W O 97/35607 PCTrUS97/04162 condition of the damaged cartilaginous tissue, the size of a wound, type of damaged tissue, the patient's age, sex, and diet, the severity of any infection, time of~1mini~tration and other clinical factors. The dosage may vary with the type of matrix used in the reconstitution and the types of additional proteins in the 5 composition. The addition of other known growth factors, such as IGF-I (insulin like growth factor I), to the final composition, may also affect the dosage.
Progress can be monitored by periodic ~ses.sment of chondrocyte survival, cartilaginous tissue formation, or cartilaginous tissue growth and/or repair. The progress can be monitored by methods known in the art, for example, X-rays, 10 arthroscopy, histomorphometric deterrninations and tetracycline labeling.
The following examples illustrate practice of the present invention in using the compositions and methods described above. The methods are useful for producing, recovering and m~inr~ining chondrocytes and/or human cartilaginous tissue inducing protein, employing the methods and compositions of cartilaginous15 tissue inducing proteins and PTHrP and equivalent PTH-like polypeptides. Although the examples demonstrate the invention with respect to BMP-2 and PTHrP1-34, withminor modifications within the skill of the art, the same results may be ~rt~in:lble with other TGF-~ proteins, particularly other BMPs, as well as equivalent PTH-like polypeptides.
EXAMPLES
1. Effect of combination of BMP and PIHrP on expression and m; -t~n~nce of cartilage specific mRNA.
PTHrP was tested for its effect on cell lines derived from E13 mouse limb 25 buds either alone or in combination with BMP-2. Cells were grown to confluence in DME medium supplemented with 10% fetal calf serum ~FCS). At confluence, cells were transferred to DME mP~ lm supplemented with 1% FCS and cultured for 1, 2, 4 or 8 days in the presence of either BMP-2 (100 ng/ml), PTHrP1-34 (100 ng/ml)(Peninsular Laboratories Inc). or a combination of BMP-2 and PTHrP1-34.
30 Northern analysis was used to determine expression of tissue specific m~NAs for cartilage (proteoglycan core protein, collagen type II and decorin), bone (osteocalcin, ~Ik~linP phosphatase and collagen type I) and hypertrophic cartilage (collagen type SlJBSTlTl)TE- SHEET (RULE 26 Wo 97/35607 PCT/USg7/04162 X). Treatment of limb bud cells with a combination of BMP-2 and PTHrP1-34 induced and m~int~inr(l expression of proteoglycan core protein and collagen type II. Expression of osteocalcin, collagen type I, alk~lin-o phosphatase and collagen - type X were all strongly inhibited by treatment with the combination of BMP-2 and PTHrP1-34, compared with treatment with BMP-2 alone. PTHrP1-34 alone did not induce expression of any of these RNAs.
These observations, combined with the reported localization of PTHrP1-34 in articular cartilage in~ tf~ that a combination of BMP-2 and PTHrP1-34 inducesthe formation and m~inten~nre of cartilaginous tissue, rather than hypertrophic cartilage or bone. This conclusion is supported by the reported localization of PTHrP in articular cartilage and the phenotype of mice cont~ining mutations to the PTHrP1-34 gene.
Example 2: Cartilage ln~ cti~n Using BMP and PIHrP
Combinations of other BMPs and PTHrP1-34 are tested for their effect on cell lines derived from E13 mouse limb buds as described in Example 1 above.
Cells are grown to confluence on medium cont~ining DME supplemented with 1%
fetal calf serum. The cells are treated with (a) BMP, such as BMP-13, alone; (b)PTHrP alone; or (c) combinations of BMP and PTHrP; in varying doses from less than 1 ng/ml up to about 5.0 x 103 ng/ml. After 10 days of treatment, histologicand Northern analyses are performed for expression of cartilage and bone markersas described in Example 1 above.
F,Y~ml)le 3: Full Thi~kn~c.c Articular C&.lilzg~ Repair Model A full thirl-nrs~ articular cartilage defect model in the femoral-patellar jointof adult rabbits is used to evaluate the ability of the combination of BMPs and PTHrP to affect cartilage and bone repair. Adult New 7e~1~nd White rabbits are anesthrti7rd and prepared for sterile surgery. A 3 x 3 mm defect through articular cartilage and into underlying subchondral bone is drilled into the patellar groove of the knee joint. The defect is either left empty, filled with collagen sponge (controls), or with collagen sponge soaked with 10 ~g rhBMP-2 alone, PTHrP alone, another BMP protein alone, or a combination of BMP and PTHrP (experimental). The SUBSTITUIE SHEET ~RULE 26) incision is closed and animals are allowed free movement within their cages for 4 wee~s. After 4 weeks the ~nim~ are hllm~n~ly e~1th~n~ti~ and the articular cartilage/subchondral bone defect is evaluated histologically for tissue architecture, quantity and quality of repair tissue. Northern analysis is performed for additional 5 phenotyping.
SUBSTlTl~E SHELT (RULE 26)
Parathyroid hormone-related peptide (PTHrP) is closely related in structure to parathyroid hormone. PTHrP is known to exist in at least three isoforms of 139, 141 and 173 amino acids. [Karaplis et al., Genes & Development, 8:277-289 (1994).
PTHrP is highly homologous to the N-terminal fragment of parathyroid hormone (PTH), and binds the same receptor as PTH. The nucleotide and amino acid sequences of the PTHrP gene from rat, mouse and human are known and may be used to produce PTHrP-like polypeptides useful in the present invention. [See Karaplis et al., Mol. Endocrin. 4:441-446 [1990][rat]; Mangin et al., PNAS
85 :597-601 (1988)[human] and Mangin et al., Gene 95:195-202 (1990)[mouse]; and Martin et al., Crit Rev Biochem Mal Biol 26:377-395 (1991).] In a preferred embodiment of the present invention, a variant of PTHrP is used in which one or more amino acids from the carboxy terminus has been deleted. For example, PTHrPl-34, which comprises the first 34 amino acids of PTHrP, is used in one preferred embodiment of the present invention. Also useful in the present invention are PTH-like polypeptides which are equivalent to PTHrPl-34 in their ability to enhance survival of chondrocytes. Such PTH-like polypeptides may include, for example, PTH, whether of human, porcine, bovine or other ~ n origin;
variants of PTH, such as those described in Wingender et al., United States Patent 5,455,329; Wingender et al., Unioed States Patent 5,457,047; and Schluter et al., United States Patent 5,457,092, and the references cited therein; as well as variants SUBSTITUTE SHEET (RULE 26) wo 97/35607 PCT/US97/04162 of the above in which one or more amino acids of PTH has been deleted from the carboxy andlor amino tennin~l portions of the molecule. The disclosures of the above publications are hereby incorporated by reference. PTH, PTHrP and the above variants may be produced via recombinant DNA engineering using the known 5 sequences of the PTH and PTHrP proteins, or may be isolated by purification.
The methods and compositions of the present invention may comprise simultaneous or sequential ~-lmini~tration of at least two active agents, a TGF~protein and a parathyroid hormone-related peptide, to a patient or site in need of cartilage repair, formation or maintenance. For sequential a~lmini~tration, one or 10 both of the active agents may be encapsulated or otherwise m~int~in~d in contact with a carrier which provides for slow release of the agent. The methods and compositions may comprise the active agents in a weight ratio of from about 90:10, to about 10:90. Preferably the active agents are present in a weight ratio of about 70:30 to 30:70. Most preferred are methods and compositions co~ lisillg the active 15 agents in a weight ratio of about 50:50.
The compositions of the invention may comprise, in addition to a TGF-,B
protein and a parathyroid hormone-related peptide, other therapentic~lly useful agents including growth factors such as epidermal growth factor (EGF), transforming growth factor-c~, activins, inhibins, platelet derived growth factor (PDGF), fibroblast 20 growth factor (FGF), and fibroblast growth factor-4 (FGF-4), parathyroid hormone (PTH), lellkt~rni~ inhibitory factor (LIF/HILDA/DIA), and insulin-like growth factors (IGF-I and IGF-II). Portions of these agents may also be used in compositions ofthe present invention. The compositions may also include an appl~l3liate matrix for in~t~nre, for supporting the composition and providing a surface for cartilage or for 25 other connective tissue growth. The matrix may provide slow release of the protein and/or the appr~liate environment for presentation thereof.
The methods and compositions of the present invention employ proteins which are able to induce cartilaginous tissue or other tissue formation in circl~Tn~t~nres where such tissue is not normally formed, and has application in the healing of 30 cartilage, for example articular cartilage tears, deformities and other cartilage defects in hllm~n~ and other animals. Such methods and compositions employing cartilaginous tissue inducing proteins may have prophylactic use in preventing SUBSTITUTE SHEET (RULE 26) damage to cartilaginous tissue, as well as use in the improved fixation of cartilage to bone or other tissues, and in repairing defects to cartilage tissue. De novo cartilaginous tissue formation induced by a composition of the present inventioncontributes to the repair of congenital, trauma in(ll~ced, or other cartilage defects of 5 other origin, and is also useful in surgery for attachment or repair of cartilage. The methods and compositions of the invention may also be useful in the treatment ofarthritis and other cartilage defects. The methods and compositions of the present invention can also be used in other indications wherein it is desirable to heal or regenerate cartilage tissue. Such indications include, without limitation, regeneration 10 or repair of injuries to the articular cartilage. The methods and compositions of the present invention may provide an environment to attract cartilage-forming cells,stim~ te growth of cartilage-forming cells or induce differentiation of progenitors of cartilage-forming cells and chondrocytes.
The compositions and methods of the present invention may also be useful 15 for treating cell populations, such as embryonic cells or stem cell populations, to enhance or enrich the growth, survival and/or differentiation of the cells into chondrocytes or other cell types. In another embodim~nt, the compositions and methods of the present invention may be used to treat chondrocytic cell lines, such as articular chondrocytes, in order to m~int~in chondrocytic phenotype and survival 20 of the cells. The treated cell populations may be useful for gene therapy applications.
The proteins useful in the methods of the present invention are useful for inducing the formation, m~ te~ re and survival of chondrocytes and/or cartilaginous tissue. It is contemplated that these proteins may have the ability to 25 induce the formation of other types of tissue, such as tendon and lig~mPnt, as well.
The cartilaginous tissue-inducing methods and compositions provided herein also may include factors encoded by the sequences similar to those of naturally-occurringTGF-,B l~lo~eins, into which modifications are naturally provided (e.g. allelic variations in the nucleotide seq-lenre which may result in amino acid changes in the 30 polypeptide) or deliberately engineered. For example, synthetic polypeptides may wholly or partially duplicate continuous sequences of the amino acid residues ofBMP-2 or BMP-13. These sequences, by virtue of sharing primary, secondary, or SUBSrlTUTE SHEET (RUL~ 26) W O 97/356D7 PCT~US97/04162 tertiary structural and conformational characteristics with cartilaginous tissue growth factor polypeptides of naturally-occurring BMP-2 or BMP-13 may possess cartilaginous or other tissue growth factor biological properties in comrnon thelcwilh. Thus, they may be employed as biologically active substitutes for 5 naturally-occurring cartilaginous tissue inducing polypeptides in therapeutic methods and compositions.
The proteins useful in the present invention, or the DNA sequences encoding therefor, may be engineered to provide one or more additional cysteine residues to increase potential dimer formation, particularly for forming dimers of the TGF-~
10 proteins. The resulting DNA sequence would be capable of producing a "cysteineadded variant" of the protein. Production of "cysteine added variants" of proteins is described in United States Patent 5,166,322, the disclosure of which is hereby incorporated by reference. Other specific mutations of the sequences of cartilaginous tissue inducing l.roteills described herein involve modifications of glycosylation sites.
15 These modifications may involve O-linked or N-linked glycosylation sites. Forin~t~nce, the absence of glycosylation or only partial glycosylation results from amino acid substitution or deletion at asparagine-linked glycosylation recognition sites. The asparagine-linked glycosylation recognition sites comprise tripeptidesequences which are specifically recognized by apprcpliate cellular glycosylation 20 enzymes. These tripeptide sequenre~ may be asparagine-X-threonine, asparagine-X-serine or asparagine-X-cysteine, where X is usually any amino acid except proline.
A variety of amino acid substitutions or deletions at one or both of the first or third amino acid positions of a glycosylation recognition site (and/or amino acid deletion at the second position) results in non-glycosylation at the modified tripeptide 25 sequence. Additionally, bacterial expression of protein will also result in production of a non-glycosylated protein, even if the glycosylation sites are left unrnodified.
The methods of the present invention for inducing cartilaginous tissue forrnation may comprise a~lministering to a patient an effective amount of a composition colll~ hlg one or more protein members of the transforming growth 30 factor-,~ (TGF-,B) ~up~lralnily together with parathyroid hormone related peptide (PTHrP), or an equivalent PTH-like polypeptide. Alternatively, the methods of the present invention for inducing cartilaginous tissue formation may comprise SU8STITUTE SHEE~ (RULE 26) W 097/3S607 PCTrUS97/04162 heterodimers comprised of two distinct monomeric units, each of which has the amino acid sequence of a member of the TGF-,B superfamily, together with PTHrP, or an equivalent PTH-like polypeptide. Preferably, the compositions of the invention comprise one or more members of the bone morphogenetic protein (BMP) family, together with PTHrP or an equivalent PTH-like polypeptide. In particular preferred embodiments, the composition comprises a bone morphogenetic protein selected from the group con~i~ting of BMP-2,BMP-4,BMP-5,BMP-7,BMP-12 and BMP-13, or heterodimers of the above, together with PTHrP or an equivalent PTH-like polypeptide. In the most preferred embo~im~n~, the composition CO~ S BMP-2, BMP-13, heterodimers of BMP-2 and BMP-13 or combinations of the above, and PTHrP or an equivalent PTH-like polypeptide. The above compositions may optionally comprise one or more additional proteins which are members of the TGF-superfamily, preferably of the BMP family of proteins. In preferred embodiments,the composition comprises BMP-2 and one or more additional proteins selected from 15 the group consisting of BMP-4, BMP-5, BMP-7, BMP-12, BMP-13 and heterodimers of the above; or BMP-13 and one or more additional proteins selected from the group con.~i~ting of BMP-2,BMP-4,BMP-5,BMP-7,BMP-12 and heterodimers of the above; each together with PTHrP or an equivalent PTH-like polypeptide. As diccllcsed above, these compositions may be used to induce the 20 formation, mai.l~enallce and survival of chondrocytes and/or cartilaginous tissue or other tissue. It is contemplated that such compositions may also be used for articular cartilage repair, wound healing and other tissue repair, such as skin repair. It is further contemplated that proteins of the invention may increase n~uro~lal survival and therefore be useful in transplantation and treatment of conditions exhibiting a decrease in neuronal survival. Compositions and methods of the invention may further be useful for induction and repair of other tissue, including wound healing.
In a preferred embodiment of the present invention, more than one protein of the TGF-~ superfamily may be used together with PTHrP to enhance the induction, maintenance and diff~nliaLion of one or more cell and/or tissue types, 30 including cartilaginous tissue. For example, a composition comprising both BMP-2 and BMP-13 implanted together with PTHrP may give rise to both bone and cartilaginous tissue. Such a composition may be useful for treating defects of the SUBSTIJUTE SI~EET (RULE 26~
W O 97135607 PCTrUS97/04162 junction between cartilage and bone, causing cartilage and bone to form simultaneously at contiguous anatomical locations, and may therefore be useful for regenerating tissue at the site of cartilage attachment to bone.
It is contemplated that the methods and compositions of the invention may also be useful in wound healing, such as skin healing and related tissue repair. The types of wounds include, but are not limited to burns, incisions and ulcers. (See, e.g. PCT Publication WO84/01106 for discussion of wound healing and related tissue repair).
The methods and compositions of the invention further comprise heteromolecules comprised of different TGF-~ moieties, in combination with PTHrPor an equivalent PTH-like polypeptide. Preferably, the heteromolecule is a heterodimer, for example, of two monomers each c-,l"L,lisillg the amino acid sequence of a BMP protein. For example, a method and composition of the invention may comprise a ~ lfi-le linked dimer COIll~l iSillg a BMP-2 protein subunit and a subunit from one of the other BMP proteins described above. Thus, in a particular embo~iment, the present invention comprises compositions and methods employing a heterodimer wherein one subunit co,~lplises the amino acid sequence of BMP-2, and one subunit comprises an amino acid sequence for a bone morphogenetic protein selected from the group consisting of BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13 or BMP-15. Further embodiments may comprise a heterodimer of other disulfide bonded TGF-,B moieties, including VGR-2, MP52, MIS, activin and TGF-,~. For example, the heterodimer may comprise one subunit comprising the amino acid sequence of BMP-2 or BMP-13, and the other subunit may comprise the amino acid of activin.
Further, compositions of the present invention may be combined with other agentsbeneficial to the treatment of the defect, wound, or tissue in question.
It is expected that the compositions and methods of the invention may act in concert with or perhaps synergistically with ~lmini~tration of other related proteins and growth factors. Further therapeutic methods and compositions of the invention therefore comprise a thelapeulic amount of at least one of the BMP proteins, whether homodimeric or heterodimeric, in combination with PTHrP or an equivalent PTH-like polypeptide, and other growth factors.
SUBSTITUTE SHEET (RULE 26) wo 97/35607 PCT/US97/04162 The preparation and formulation of such physiologically acceptable protein compositions, having due regard to pH, isotonicity, stability and the like, is within the skill of the art. The therapeutic compositions are also presently valuable for veterinary applications due to the lack of species specificity in TGF-~ proteins.
5 Particularly domestic animals and thoroughbred horses in addition to humans are desired patients for such treatment with the compositions of the present invention.
The therapeutic method includes :~mini~tering the composition topically, systemically, or locally as an injectable and/or implant or device. When admin-istered, the therapeutic composition for use in this invention is, of course, in a 10 pyrogen-free, physiologically acceptable form. Further, the composition may desirably be encapsulated or injected in a viscous form for delivery to the site of tissue damage. Topical a(lmini~tration may be suitable for wound healing and tissue repair. Therapeutic~lly useful agents other than the proteins which may also optionally be included in the composition as described above, may alternatively or 15 additionally, be ~lmini~tered .siml~lt:~neously or sequentially with the composition in the methods of the invention. In addition, the compositions of the present invention may be used in conjunction with presently available treatments for cartilage injuries, such as suture (e.g., vicryl sutures or surgical gut sutures, Ethicon Inc., Somerville, NJ) or cartilage allograft or autograft, in order to enh~nre or accelerate 20 the healing potential of the suture or graft. For example, the suture, allograft or autograft may be soaked in the compositions of the present invention prior to implantation. It may also be possible to incorporate the protein or composition of the invention onto suture materials, for example, by freeze-drying.
The compositions of the present invention may include an app~ul)l iate matrix 25 and/or sequestering agent as a carrier. For in~t~nre, the matrix may support the composition or provide a surface for cartilaginous tissue formation and/or othertissue formation. The matrix may provide slow release of the protein and/or the ap~lopliate environment for presentation thereof. The sequestering agent may be a substance which aids in ease of a(lmini~tration through injection or other means, 30 or may slow the migration of protein from the site of application.
The choice of a carrier material is based on biocompatibility, biodegradability, mechanical properties, cosmetic appearance and interface SUBSTITIJTE SHEET (RULE 26) W 097/35607 PCT~US97/04162 properties. The particular application of the compositions will define the ~yplopfiate formulation. Potential matrices for the compositions may be biodegradable and chemically defined. Further matrices are comprised of pure proteins or extracellular matrix components. Other potential matrices are nonbiodegradable and chemically 5 defined. Preferred matrices include collagen-based materials, including sponges, such as Helistat~ (Integra LifeSciences, Plainsboro, N.J.), orcollagen in an injectable form, as well as sequestering agents, which may be biodegradable, for example hyaluronic acid derived. Biodegradable materials, such as cellulose films, or surgical meshes, may also serve as matrices. Such materials could be sutured into 10 an injury site, or wrapped around the cartilage.
Another preferred class of carrier are polymeric matrices, including polymers of poly(lactic acid), poly(glycolic acid) and copolymers of lactic acid and glycolic acid. These matrices may be in the form of a sponge, or in the form of porous particles, and may also include a sequçstering agent. Suitable polymer matrices are described, for example, in WO93/00050, the disclosure of which is incorporated herein by reference.
Preferred f~milil~s of se~uestering agents include blood, fibrin clot and/or cellulosic materials such as alkylcelluloses (including hydroxyalkylcelluloses),including methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl-methylcellulose, and carboxymethylcellulose, the most preferred being cationic salts of carboxymethylcellulose (CMC). Other preferred sequestering agents include hyaluronic acid, sodium alginate, poly(ethylene glycol), polyoxyethylene oxide, carboxyvinyl polymer and poly(vinyl alcohol). The amount of seql~estering agent useful herein is 0.5-20 wt%, preferably 1-10 wt%
based on total formulation weight, which represents the amount n~ces~ry to prevent desorbtion of the protein from the polymer matrix and to provide a~ opliate h~n-lling of the composition, yet not so much that the progenitor cells are prevented from infiltrating the matrix, thereby providing the protein the opportunity to assist the activity of the progenitor cells.
Additional optional components useful in the practice of the subject application include, e.g. cryogenic protectors such as m~nnitol, sucrose, lactose, glucose, or glycine (to protect the protein from degradation during Iyophilization), SUBSTITUTE SHEET (RULE 26) W O 97/35607 PCTrUS97/04162 antimicrobial preservatives such as methyl and propyl parabens and benzyl alcohol;
antioxidants such as EDTA, citrate and BHT (butylated hydroxytoluene); and surfactants such as poly(sorbates) and poly(oxyethylenes); etc.
As described above, the compositions of the invention may be employed in 5 methods for treating a number of cartilage defects, such as the regeneration of cartilaginous tissue in areas of cartilage damage, to assist in repair of tears of cartilage tissue, and various other types of tissue defects or wounds. These methods, according to the invention, entail ~Amini~tering to a patient nee(ling such cartilaginous tissue or other tissue repair, a composition comprising an effective 10 amount of an osteogenic or cartilage-inducing member of the transforming factor beta [TGF-~] superfamily of proteins, such as a bone morphogenetic protein [BMP], preferably BMP-2,BMP-4,BMP-S,BMP-7,BMP-12 and BMP-13, in combination with parathyroid hormone-related peptide [PTHrP] or an equivalent PTH-like polypeptide.
In another embodiment, the methods may entail ~lminictration of a heterodimeric protein in which each of the monomers cu~ lises the amino acid sequence of an osteogenic or cartilage-inducing member of the TGF,~ superfamily of proteins, such as a BMP, particularly, BMP-2,BMP-4,BMP-5,BMP-7,BMP-12 and BMP-13, in combination with PTHrP or an equivalent PTHrP-like polypeptide.
In a preferred embodiment, each of the monomers is selected from the group consisting of BMP-2,BMP-4,BMP-5,BMP-7,BMP-12 and BMP-13.
Thus, a further aspect of the invention is a therapeutic method and composition for inducing or m~int~ining chondrocytes or cartilaginous tissue, for repairing cartilaginous tissue, for repairing cartilage as well as treating arthritis and other conditions related to arthritis or cartilage defects. Such compositions comprise a therape~tir~lly effective amount of one or more osteogenic or cartilaginous tissue inducing proteins, such as BMP-2 or BMP-13, in combination with PTHrP or an equivalent PTH-like polypeptide, in admixture with a pharm~relltic~lly acceptable vehicle, carrier or matrix.
The dosage regimen will be det~rminrd by the attending physician considering various factors which modify the action of the composition, e.g., amount of cartilaginous tissue desired to be formed, the site of cartilaginous tissue damage, the SVBSrlTUTE SHEET (RUlE 26) W O 97/35607 PCTrUS97/04162 condition of the damaged cartilaginous tissue, the size of a wound, type of damaged tissue, the patient's age, sex, and diet, the severity of any infection, time of~1mini~tration and other clinical factors. The dosage may vary with the type of matrix used in the reconstitution and the types of additional proteins in the 5 composition. The addition of other known growth factors, such as IGF-I (insulin like growth factor I), to the final composition, may also affect the dosage.
Progress can be monitored by periodic ~ses.sment of chondrocyte survival, cartilaginous tissue formation, or cartilaginous tissue growth and/or repair. The progress can be monitored by methods known in the art, for example, X-rays, 10 arthroscopy, histomorphometric deterrninations and tetracycline labeling.
The following examples illustrate practice of the present invention in using the compositions and methods described above. The methods are useful for producing, recovering and m~inr~ining chondrocytes and/or human cartilaginous tissue inducing protein, employing the methods and compositions of cartilaginous15 tissue inducing proteins and PTHrP and equivalent PTH-like polypeptides. Although the examples demonstrate the invention with respect to BMP-2 and PTHrP1-34, withminor modifications within the skill of the art, the same results may be ~rt~in:lble with other TGF-~ proteins, particularly other BMPs, as well as equivalent PTH-like polypeptides.
EXAMPLES
1. Effect of combination of BMP and PIHrP on expression and m; -t~n~nce of cartilage specific mRNA.
PTHrP was tested for its effect on cell lines derived from E13 mouse limb 25 buds either alone or in combination with BMP-2. Cells were grown to confluence in DME medium supplemented with 10% fetal calf serum ~FCS). At confluence, cells were transferred to DME mP~ lm supplemented with 1% FCS and cultured for 1, 2, 4 or 8 days in the presence of either BMP-2 (100 ng/ml), PTHrP1-34 (100 ng/ml)(Peninsular Laboratories Inc). or a combination of BMP-2 and PTHrP1-34.
30 Northern analysis was used to determine expression of tissue specific m~NAs for cartilage (proteoglycan core protein, collagen type II and decorin), bone (osteocalcin, ~Ik~linP phosphatase and collagen type I) and hypertrophic cartilage (collagen type SlJBSTlTl)TE- SHEET (RULE 26 Wo 97/35607 PCT/USg7/04162 X). Treatment of limb bud cells with a combination of BMP-2 and PTHrP1-34 induced and m~int~inr(l expression of proteoglycan core protein and collagen type II. Expression of osteocalcin, collagen type I, alk~lin-o phosphatase and collagen - type X were all strongly inhibited by treatment with the combination of BMP-2 and PTHrP1-34, compared with treatment with BMP-2 alone. PTHrP1-34 alone did not induce expression of any of these RNAs.
These observations, combined with the reported localization of PTHrP1-34 in articular cartilage in~ tf~ that a combination of BMP-2 and PTHrP1-34 inducesthe formation and m~inten~nre of cartilaginous tissue, rather than hypertrophic cartilage or bone. This conclusion is supported by the reported localization of PTHrP in articular cartilage and the phenotype of mice cont~ining mutations to the PTHrP1-34 gene.
Example 2: Cartilage ln~ cti~n Using BMP and PIHrP
Combinations of other BMPs and PTHrP1-34 are tested for their effect on cell lines derived from E13 mouse limb buds as described in Example 1 above.
Cells are grown to confluence on medium cont~ining DME supplemented with 1%
fetal calf serum. The cells are treated with (a) BMP, such as BMP-13, alone; (b)PTHrP alone; or (c) combinations of BMP and PTHrP; in varying doses from less than 1 ng/ml up to about 5.0 x 103 ng/ml. After 10 days of treatment, histologicand Northern analyses are performed for expression of cartilage and bone markersas described in Example 1 above.
F,Y~ml)le 3: Full Thi~kn~c.c Articular C&.lilzg~ Repair Model A full thirl-nrs~ articular cartilage defect model in the femoral-patellar jointof adult rabbits is used to evaluate the ability of the combination of BMPs and PTHrP to affect cartilage and bone repair. Adult New 7e~1~nd White rabbits are anesthrti7rd and prepared for sterile surgery. A 3 x 3 mm defect through articular cartilage and into underlying subchondral bone is drilled into the patellar groove of the knee joint. The defect is either left empty, filled with collagen sponge (controls), or with collagen sponge soaked with 10 ~g rhBMP-2 alone, PTHrP alone, another BMP protein alone, or a combination of BMP and PTHrP (experimental). The SUBSTITUIE SHEET ~RULE 26) incision is closed and animals are allowed free movement within their cages for 4 wee~s. After 4 weeks the ~nim~ are hllm~n~ly e~1th~n~ti~ and the articular cartilage/subchondral bone defect is evaluated histologically for tissue architecture, quantity and quality of repair tissue. Northern analysis is performed for additional 5 phenotyping.
SUBSTlTl~E SHELT (RULE 26)
Claims (15)
1, A composition comprising:
(a) at least one protein member of the bone morphogenetic protein (BMP) family of proteins; and (b) parathyroid hormone related peptide (PTHrP).
(a) at least one protein member of the bone morphogenetic protein (BMP) family of proteins; and (b) parathyroid hormone related peptide (PTHrP).
2. The composition of claim 1, wherein the bone morphogenetic protein is selected from the group consisting of BMP-2, BMP-4, BMP-5, BMP-7, BMP-12 and BMP-13.
3. The composition of claim 1, wherein the bone morphogenetic protein is BMP-2.
4. The composition of claim 2, wherein the bone morphogenetic protein is BMP-13.
5. The composition of claim 3, wherein said composition further comprises one or more additional proteins which are members of the BMP subfamily of proteins.
6. The composition of claim 5, wherein the additional proteins are selected from the group consisting of BMP-4, BMP-5, BMP-7, BMP-12 and BMP-13.
7. The composition of claim 4, wherein said composition further comprises one or more additional proteins which are members of the BMP subfamily of proteins.
8. The composition of claim 7, wherein the additional proteins are selected from the group consisting of BMP-2, BMP-4, BMP-5, BMP-7 and BMP-12.
9. A composition comprising:
(a) at least one protein member of the transforming growth factor-.beta. (TGF-.beta.) superfamily selected from the group consisting of BMP-2, BMP-13, heterodimers ofBMP-2 and BMP-13 and combinations thereof; and (b) parathyroid hormone related peptide (PTHrP).
(a) at least one protein member of the transforming growth factor-.beta. (TGF-.beta.) superfamily selected from the group consisting of BMP-2, BMP-13, heterodimers ofBMP-2 and BMP-13 and combinations thereof; and (b) parathyroid hormone related peptide (PTHrP).
10. A composition comprising:
(a) at least one protein member of the transforming growth factor-.beta. (TGF-.beta.) superfamily selected from the group consisting of BMP-2, BMP-13, heterodimers ofBMP-2 and BMP-13 and combinations thereof; and (b) a truncated parathyroid hormone related peptide (PTHrP) comprising amino acids 1 to 34 of PTHrP.
(b) a truncated parathyroid hormone related peptide (PTHrP) comprising amino acids 1 to 34 of PTHrP.
(a) at least one protein member of the transforming growth factor-.beta. (TGF-.beta.) superfamily selected from the group consisting of BMP-2, BMP-13, heterodimers ofBMP-2 and BMP-13 and combinations thereof; and (b) a truncated parathyroid hormone related peptide (PTHrP) comprising amino acids 1 to 34 of PTHrP.
(b) a truncated parathyroid hormone related peptide (PTHrP) comprising amino acids 1 to 34 of PTHrP.
11. The composition of claim 10 wherein the TGF-.beta. superfamily member is BMP-2.
12. The composition of claim 10 wherein the TGF-.beta. superfamily member is BMP-13.
13. A method for inducing the formation or maintenance of cartilaginous tissue in a patient, comprising administering to the patient an effective amount of the composition of claim 1.
14. A method according to claim 13, wherein the patient is suffering from arthritis.
15. A method according to claim 13, wherein the patient is suffering from an articular cartilage defect or damage.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US08/622,101 | 1996-03-26 | ||
| US08/622,101 US5700774A (en) | 1996-03-26 | 1996-03-26 | Compositions comprising bone morphogenic proteins and truncated parathyroid hormone related peptide, and methods of inducing cartilage by administration of same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2249212A1 true CA2249212A1 (en) | 1997-10-02 |
Family
ID=24492945
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002249212A Abandoned CA2249212A1 (en) | 1996-03-26 | 1997-03-17 | Methods of tissue induction using a combination of bone morphogenetic protein and parathyroid hormone-related peptide |
Country Status (12)
| Country | Link |
|---|---|
| US (2) | US5700774A (en) |
| EP (1) | EP0906120B1 (en) |
| JP (1) | JP2000507944A (en) |
| KR (1) | KR19990087489A (en) |
| AT (1) | ATE219685T1 (en) |
| AU (1) | AU727312B2 (en) |
| CA (1) | CA2249212A1 (en) |
| DE (1) | DE69713597T2 (en) |
| FI (1) | FI981916A (en) |
| NO (1) | NO984345L (en) |
| OA (1) | OA10885A (en) |
| WO (1) | WO1997035607A1 (en) |
Families Citing this family (80)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ATE238417T1 (en) | 1991-11-04 | 2003-05-15 | Inst Genetics Llc | RECOMBINANT BONE MORPHOGENETIC PROTEIN HETERODIMERS, COMPOSITIONS AND METHODS OF USE |
| US5942496A (en) | 1994-02-18 | 1999-08-24 | The Regent Of The University Of Michigan | Methods and compositions for multiple gene transfer into bone cells |
| US6551618B2 (en) | 1994-03-15 | 2003-04-22 | University Of Birmingham | Compositions and methods for delivery of agents for neuronal regeneration and survival |
| US5906827A (en) * | 1994-06-03 | 1999-05-25 | Creative Biomolecules, Inc. | Matrix for the manufacture of autogenous replacement body parts |
| GB9721797D0 (en) * | 1997-10-14 | 1997-12-17 | Univ Cardiff | Materials and methods relating to cartilage repair |
| AU752816B2 (en) * | 1997-11-28 | 2002-10-03 | Curis, Inc. | An active hedgehog-protein-mutant, a process for its preparation and its use for pharmaceutical purposes |
| US6897297B1 (en) * | 1997-12-03 | 2005-05-24 | Curis, Inc. | Hydrophobically-modified protein compositions and methods |
| US6171340B1 (en) | 1998-02-27 | 2001-01-09 | Mcdowell Charles L. | Method and device for regenerating cartilage in articulating joints |
| DK0953576T3 (en) * | 1998-04-30 | 2006-03-20 | Curis Inc | Active hedgehog protein conjugate, process for its preparation and use |
| AU756785B2 (en) | 1998-07-13 | 2003-01-23 | University Of Southern California | Methods for accelerating bone and cartilage growth and repair |
| US6916783B2 (en) * | 1998-07-13 | 2005-07-12 | University Of Southern California | Methods for accelerating bone and cartilage growth and repair |
| US7067144B2 (en) * | 1998-10-20 | 2006-06-27 | Omeros Corporation | Compositions and methods for systemic inhibition of cartilage degradation |
| US6727224B1 (en) * | 1999-02-01 | 2004-04-27 | Genetics Institute, Llc. | Methods and compositions for healing and repair of articular cartilage |
| US6197061B1 (en) * | 1999-03-01 | 2001-03-06 | Koichi Masuda | In vitro production of transplantable cartilage tissue cohesive cartilage produced thereby, and method for the surgical repair of cartilage damage |
| EP2305324B1 (en) | 1999-03-25 | 2014-09-17 | Metabolix, Inc. | Medical devices and applications of polyhydroxyalkanoate polymers |
| US6315992B1 (en) | 1999-06-30 | 2001-11-13 | Tissuegene Co. | Generating cartilage in a mammal using fibroblasts transfected with a vector encoding TGF-β-1 |
| US7338655B1 (en) | 1999-06-30 | 2008-03-04 | Tissuegene, Inc. | Gene therapy using TGF-β |
| CA2378880C (en) * | 1999-07-21 | 2010-02-23 | Omeros Medical Systems, Inc. | Solutions and methods for inhibition of pain, inflammation and cartilage degradation |
| WO2006089023A2 (en) | 2005-02-16 | 2006-08-24 | The Regents Of The University Of California | Pharmaceutical compositions for treating or preventing bone conditions |
| US7687462B2 (en) * | 1999-10-05 | 2010-03-30 | The Regents Of The University Of California | Composition for promoting cartilage formation or repair comprising a nell gene product and method of treating cartilage-related conditions using such composition |
| US6770078B2 (en) | 2000-01-14 | 2004-08-03 | Peter M. Bonutti | Movable knee implant and methods therefor |
| US7635390B1 (en) | 2000-01-14 | 2009-12-22 | Marctec, Llc | Joint replacement component having a modular articulating surface |
| US6702821B2 (en) * | 2000-01-14 | 2004-03-09 | The Bonutti 2003 Trust A | Instrumentation for minimally invasive joint replacement and methods for using same |
| AU2001278742A1 (en) * | 2000-08-16 | 2002-02-25 | Chugai Seiyaku Kabushiki Kaisha | Agents for ameliorating symptoms caused by joint diseases |
| US7708741B1 (en) | 2001-08-28 | 2010-05-04 | Marctec, Llc | Method of preparing bones for knee replacement surgery |
| JP2005506860A (en) | 2001-09-15 | 2005-03-10 | ラツシユ−プレスビテリアン−セント・リユークズ・メデイカル・センター | Layered cartilage tissue and engineering method thereof |
| WO2003028655A2 (en) * | 2001-10-04 | 2003-04-10 | Massachusetts Institute Of Technology | Effect of bone morphogenetic proteins on engineered cartilage |
| US20030165473A1 (en) * | 2001-11-09 | 2003-09-04 | Rush-Presbyterian-St. Luke's Medical Center | Engineered intervertebral disc tissue |
| US7015195B2 (en) * | 2002-01-10 | 2006-03-21 | Osteotrophin, Llc | Treatment of bone disorders with skeletal anabolic drugs |
| CN1653179B (en) * | 2002-03-12 | 2012-07-25 | 组织基因股份有限公司 | Cartilage regeneration using chondrocyte and TGF-beta |
| US7431922B2 (en) * | 2002-03-29 | 2008-10-07 | Tissuegene, Inc. | Bioadhesive directed somatic cell therapy |
| US7005127B2 (en) * | 2002-03-29 | 2006-02-28 | Tissuegene, Inc. | Mixed-cell gene therapy |
| CA2490409A1 (en) | 2002-06-28 | 2004-01-08 | Centocor, Inc. | Mammalian ch1 deleted mimetibodies, compositions, methods and uses |
| EP1542711A4 (en) * | 2002-08-13 | 2009-07-01 | Wyeth Corp | PEPTIDES AS SOLUBILIZING EXCIPIENTS FOR TRANSFORMING GROWTH FACTOR s PROTEINS |
| US20050209142A1 (en) * | 2002-11-20 | 2005-09-22 | Goran Bertilsson | Compounds and methods for increasing neurogenesis |
| US8551100B2 (en) | 2003-01-15 | 2013-10-08 | Biomet Manufacturing, Llc | Instrumentation for knee resection |
| US7789885B2 (en) | 2003-01-15 | 2010-09-07 | Biomet Manufacturing Corp. | Instrumentation for knee resection |
| US7837690B2 (en) | 2003-01-15 | 2010-11-23 | Biomet Manufacturing Corp. | Method and apparatus for less invasive knee resection |
| US7887542B2 (en) | 2003-01-15 | 2011-02-15 | Biomet Manufacturing Corp. | Method and apparatus for less invasive knee resection |
| US20080194472A1 (en) * | 2003-03-27 | 2008-08-14 | Jeffrey Allen Whitsett | Use of Fgf-18 Protein, Target Proteins and Their Respective Encoding Nucleotide Sequences to Induce Cartilage Formation |
| US7067123B2 (en) | 2003-04-29 | 2006-06-27 | Musculoskeletal Transplant Foundation | Glue for cartilage repair |
| US7488348B2 (en) | 2003-05-16 | 2009-02-10 | Musculoskeletal Transplant Foundation | Cartilage allograft plug |
| US7901457B2 (en) | 2003-05-16 | 2011-03-08 | Musculoskeletal Transplant Foundation | Cartilage allograft plug |
| EP1675608B1 (en) | 2003-09-12 | 2007-03-21 | Wyeth | Injectable calcium phosphate solid rods for delivery of osteogenic proteins |
| US7488324B1 (en) | 2003-12-08 | 2009-02-10 | Biomet Manufacturing Corporation | Femoral guide for implanting a femoral knee prosthesis |
| US8697139B2 (en) | 2004-09-21 | 2014-04-15 | Frank M. Phillips | Method of intervertebral disc treatment using articular chondrocyte cells |
| US7837740B2 (en) | 2007-01-24 | 2010-11-23 | Musculoskeletal Transplant Foundation | Two piece cancellous construct for cartilage repair |
| US20060166251A1 (en) * | 2005-01-26 | 2006-07-27 | Archambault Joanne M | Use of sFRPs as markers of BMP activity |
| US7695479B1 (en) | 2005-04-12 | 2010-04-13 | Biomet Manufacturing Corp. | Femoral sizer |
| US7815926B2 (en) | 2005-07-11 | 2010-10-19 | Musculoskeletal Transplant Foundation | Implant for articular cartilage repair |
| US7622438B1 (en) | 2005-07-18 | 2009-11-24 | Acologix, Inc. | Protein formulation for promoting hard tissue formation |
| AU2006292224B2 (en) | 2005-09-19 | 2013-08-01 | Histogenics Corporation | Cell-support matrix and a method for preparation thereof |
| US8070752B2 (en) | 2006-02-27 | 2011-12-06 | Biomet Manufacturing Corp. | Patient specific alignment guide and inter-operative adjustment |
| US9173661B2 (en) | 2006-02-27 | 2015-11-03 | Biomet Manufacturing, Llc | Patient specific alignment guide with cutting surface and laser indicator |
| US8603180B2 (en) | 2006-02-27 | 2013-12-10 | Biomet Manufacturing, Llc | Patient-specific acetabular alignment guides |
| US10278711B2 (en) | 2006-02-27 | 2019-05-07 | Biomet Manufacturing, Llc | Patient-specific femoral guide |
| US9345548B2 (en) | 2006-02-27 | 2016-05-24 | Biomet Manufacturing, Llc | Patient-specific pre-operative planning |
| US8591516B2 (en) | 2006-02-27 | 2013-11-26 | Biomet Manufacturing, Llc | Patient-specific orthopedic instruments |
| US9289253B2 (en) | 2006-02-27 | 2016-03-22 | Biomet Manufacturing, Llc | Patient-specific shoulder guide |
| US20150335438A1 (en) | 2006-02-27 | 2015-11-26 | Biomet Manufacturing, Llc. | Patient-specific augments |
| US9918740B2 (en) | 2006-02-27 | 2018-03-20 | Biomet Manufacturing, Llc | Backup surgical instrument system and method |
| US9113971B2 (en) | 2006-02-27 | 2015-08-25 | Biomet Manufacturing, Llc | Femoral acetabular impingement guide |
| US8407067B2 (en) | 2007-04-17 | 2013-03-26 | Biomet Manufacturing Corp. | Method and apparatus for manufacturing an implant |
| US7780672B2 (en) | 2006-02-27 | 2010-08-24 | Biomet Manufacturing Corp. | Femoral adjustment device and associated method |
| US9339278B2 (en) | 2006-02-27 | 2016-05-17 | Biomet Manufacturing, Llc | Patient-specific acetabular guides and associated instruments |
| US9907659B2 (en) | 2007-04-17 | 2018-03-06 | Biomet Manufacturing, Llc | Method and apparatus for manufacturing an implant |
| US7695520B2 (en) | 2006-05-31 | 2010-04-13 | Biomet Manufacturing Corp. | Prosthesis and implementation system |
| US9795399B2 (en) | 2006-06-09 | 2017-10-24 | Biomet Manufacturing, Llc | Patient-specific knee alignment guide and associated method |
| EP2117578B1 (en) * | 2007-01-22 | 2015-04-22 | OrthoTrophix, Inc. | A peptide composition and a method of promoting cartilage formation |
| US8435551B2 (en) | 2007-03-06 | 2013-05-07 | Musculoskeletal Transplant Foundation | Cancellous construct with support ring for repair of osteochondral defects |
| CA2717725A1 (en) | 2008-03-05 | 2009-09-11 | Musculoskeletal Transplant Foundation | Cancellous constructs, cartilage particles and combinations of cancellous constructs and cartilage particles |
| US8513193B2 (en) | 2008-10-13 | 2013-08-20 | University Of Rochester | Protecting and repairing cartilage and musculoskeletal soft tissues |
| CN102292092B (en) | 2008-11-25 | 2014-05-07 | 组织基因公司 | Primed cell therapy |
| US8530420B2 (en) * | 2008-12-16 | 2013-09-10 | Kaohsiung Medical University | Treatment of arthritis with parathyroid hormone |
| US9968376B2 (en) | 2010-11-29 | 2018-05-15 | Biomet Manufacturing, Llc | Patient-specific orthopedic instruments |
| US9241745B2 (en) | 2011-03-07 | 2016-01-26 | Biomet Manufacturing, Llc | Patient-specific femoral version guide |
| EP2875125B1 (en) * | 2012-07-20 | 2018-10-24 | Alfred C. Kuo | Methods for producing cartilage and bone |
| US10077420B2 (en) | 2014-12-02 | 2018-09-18 | Histogenics Corporation | Cell and tissue culture container |
| US20190381341A1 (en) * | 2017-02-01 | 2019-12-19 | The Johns Hopkins University | Parathyroid hormone and regeneration of degenerative discs disease |
| US10722310B2 (en) | 2017-03-13 | 2020-07-28 | Zimmer Biomet CMF and Thoracic, LLC | Virtual surgery planning system and method |
Family Cites Families (26)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5459047A (en) * | 1986-07-01 | 1995-10-17 | Genetics Institute, Inc. | BMP-6 proteins |
| US5187076A (en) * | 1986-07-01 | 1993-02-16 | Genetics Institute, Inc. | DNA sequences encoding BMP-6 proteins |
| US5013649A (en) * | 1986-07-01 | 1991-05-07 | Genetics Institute, Inc. | DNA sequences encoding osteoinductive products |
| US5543394A (en) * | 1986-07-01 | 1996-08-06 | Genetics Institute, Inc. | Bone morphogenetic protein 5(BMP-5) compositions |
| ZA874681B (en) * | 1986-07-01 | 1988-04-27 | Genetics Inst | Novel osteoinductive factors |
| US5106748A (en) * | 1986-07-01 | 1992-04-21 | Genetics Institute, Inc. | Dna sequences encoding 5 proteins |
| US5108922A (en) * | 1986-07-01 | 1992-04-28 | Genetics Institute, Inc. | DNA sequences encoding BMP-1 products |
| US5457092A (en) * | 1987-07-30 | 1995-10-10 | Gesellschaft Fur Biotechnologische Forschung Mbh (Gbf) | Methods of promoting bone growth in mammals comprising administration of modified parathyroid hormone |
| JPH03504201A (en) * | 1989-02-23 | 1991-09-19 | ゲゼルシャフト・フュア・ビオテクノロギッシェ・フォルシュンク・ミット・ベシュレンクテル・ハフツング （ゲー・ベー・エフ） | DNA sequences encoding PTH variants, PTH variants, expression vectors, bacterial hosts, compositions for use and treatment |
| ATE209251T1 (en) * | 1990-05-16 | 2001-12-15 | Genetics Inst | PROTEINS PRODUCING BONE AND CARTILAGE FORMATION |
| US5364839A (en) * | 1990-06-18 | 1994-11-15 | Genetics Institute, Inc. | Osteoinductive pharmaceutical formulations |
| US5118667A (en) * | 1991-05-03 | 1992-06-02 | Celtrix Pharmaceuticals, Inc. | Bone growth factors and inhibitors of bone resorption for promoting bone formation |
| DE69213739T2 (en) * | 1991-06-21 | 1997-02-20 | Genetics Inst | MEDICINAL PRODUCTS CONTAINING OSTEOGENIC PROTEINS |
| KR100255415B1 (en) * | 1991-06-25 | 2000-05-01 | 브루스 엠. 에이센 | Bone morphogenetic protein-9 compositions |
| ATE238417T1 (en) * | 1991-11-04 | 2003-05-15 | Inst Genetics Llc | RECOMBINANT BONE MORPHOGENETIC PROTEIN HETERODIMERS, COMPOSITIONS AND METHODS OF USE |
| HU218845B (en) * | 1992-02-12 | 2000-12-28 | Biopharm Gesellschaft Zur Biotechnologischen Entwicklung Von Pharmaka Mbh. | Dna sequences encoding novel growth/differentiation factors, pharmaceutical preparations containing said factors, dna molecules, mosts, antibodies and diagnostic methods |
| IL106278A0 (en) * | 1992-07-13 | 1993-11-15 | Sumitomo Metal Ind | Bone formation-inducing protein |
| JPH08505771A (en) * | 1993-01-12 | 1996-06-25 | ジョーンズ ホプキンス ユニバーシティー スクール オブ メディシン | Growth differentiation factor-3 |
| EP1439190A1 (en) * | 1993-01-12 | 2004-07-21 | The Johns Hopkins University School Of Medicine | Growth differentiation factor-5 |
| WO1994015966A1 (en) * | 1993-01-12 | 1994-07-21 | Johns Hopkins University School Of Medicine | Growth differentiation factor-9 |
| JPH09507829A (en) * | 1993-03-19 | 1997-08-12 | ジョーンズ ホプキンス ユニバーシティー スクール オブ メディシン | Growth differentiation factor-8 |
| KR100231640B1 (en) * | 1993-05-12 | 1999-12-01 | 토마스 제이 데스로저 | Bmp-10 compositions |
| EP2272959A1 (en) * | 1993-05-12 | 2011-01-12 | Genetics Institute, LLC | BMP-11 compositions |
| WO1995001801A1 (en) * | 1993-07-09 | 1995-01-19 | The Johns Hopkins University School Of Medicine | Growth differentiation factor-6 |
| JPH09503904A (en) * | 1993-07-09 | 1997-04-22 | ザ ジョーンズ ホプキンス ユニバーシティー スクール オブ メディシン | Growth differentiation factor-7 |
| CA2176942C (en) * | 1993-12-07 | 2011-11-01 | Anthony J. Celeste | Bmp-12, bmp-13 and tendon-inducing compositions thereof |
-
1996
- 1996-03-26 US US08/622,101 patent/US5700774A/en not_active Expired - Lifetime
-
1997
- 1997-03-17 WO PCT/US1997/004162 patent/WO1997035607A1/en not_active Application Discontinuation
- 1997-03-17 AU AU23290/97A patent/AU727312B2/en not_active Ceased
- 1997-03-17 CA CA002249212A patent/CA2249212A1/en not_active Abandoned
- 1997-03-17 KR KR1019980706917A patent/KR19990087489A/en not_active Application Discontinuation
- 1997-03-17 DE DE69713597T patent/DE69713597T2/en not_active Expired - Lifetime
- 1997-03-17 AT AT97916010T patent/ATE219685T1/en active
- 1997-03-17 EP EP97916010A patent/EP0906120B1/en not_active Expired - Lifetime
- 1997-03-17 JP JP9534452A patent/JP2000507944A/en active Pending
- 1997-09-10 US US08/926,942 patent/US5846931A/en not_active Expired - Lifetime
-
1998
- 1998-09-08 FI FI981916A patent/FI981916A/en not_active IP Right Cessation
- 1998-09-18 NO NO984345A patent/NO984345L/en not_active Application Discontinuation
- 1998-09-25 OA OA9800179A patent/OA10885A/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| AU2329097A (en) | 1997-10-17 |
| WO1997035607A1 (en) | 1997-10-02 |
| JP2000507944A (en) | 2000-06-27 |
| OA10885A (en) | 2003-02-18 |
| FI981916A0 (en) | 1998-09-08 |
| ATE219685T1 (en) | 2002-07-15 |
| DE69713597D1 (en) | 2002-08-01 |
| EP0906120A1 (en) | 1999-04-07 |
| KR19990087489A (en) | 1999-12-27 |
| NO984345D0 (en) | 1998-09-18 |
| US5846931A (en) | 1998-12-08 |
| EP0906120B1 (en) | 2002-06-26 |
| FI981916A (en) | 1998-09-08 |
| AU727312B2 (en) | 2000-12-07 |
| NO984345L (en) | 1998-09-18 |
| DE69713597T2 (en) | 2003-02-20 |
| US5700774A (en) | 1997-12-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5846931A (en) | Compositions comprising bone morphogenic proteins and truncated parathyroid hormone related peptide and methods of inducing cartilage by administration of same | |
| US5902785A (en) | Cartilage induction by bone morphogenetic proteins | |
| US5604204A (en) | Method of inducing bone growth using TGF-β | |
| Chubinskaya et al. | Regulation of osteogenic proteins by chondrocytes | |
| Kirker-Head | Potential applications and delivery strategies for bone morphogenetic proteins | |
| US5364839A (en) | Osteoinductive pharmaceutical formulations | |
| EP0679097B1 (en) | Tgf-beta formulation for inducing bone growth | |
| US5422340A (en) | TGF-βformulation for inducing bone growth | |
| EP0957943B2 (en) | Matrix-free osteogenic devices, implants and methods of use thereof | |
| AU772479B2 (en) | Repair of larynx, trachea, and other fibrocartilaginous tissues | |
| Vukičević et al. | Discovery and clinical applications of bone morphogenetic proteins | |
| US20040176287A1 (en) | Bmp binding proteins for use in bone or cartilage regeneration | |
| DE60123218T2 (en) | PRODUCT FOR BIOLOGICAL BIND WEB ANCHORING OF BONE | |
| KR20000036012A (en) | Bone morphogenetic protein-16 (bmp-16) compositions | |
| US20050287135A1 (en) | Injectable solid hyaluronic acid carriers for delivery of osteogenic proteins | |
| Bostrom et al. | Biologics in bone healing | |
| McKinney et al. | A Bone Regeneration Study: Transforming Growth Factor-β: 1: and Its Delivery | |
| NZ248446A (en) | Bone growth composition containing transforming growth factor beta (tgf-beta) and demineralised bone | |
| Chiba et al. | Histological study of effect of bone morphogenetic protein derived from bovine tooth on periosteum in rats | |
| AU2002302768A1 (en) | BMP binding proteins for use in bone or cartilage regeneration |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Discontinued |